My RNA is already enriched for miRNAs, how much should I use in the real-time PCR experiments?
The miRCURY LNA™ microRNA PCR system is developed for use on total RNA and we do not recommend enriching for small RNAs. Samples of enriched microRNAs are difficult to quantitate accurately making it very tricky to ensure the same amount of sample is added to each reaction. If necessary, a total RNA equivalent should be used for the enriched sample, e.g. use a proportional amount of enriched sample resulting from 1-10 ng of total RNA. It may be necessary to try a couple of different amounts of enriched sample to ensure that the results fall within the linear range of the assay.
